螨变应原点刺制品生物活性测定方法

A method for determination of bioactivity of mite allergen prick solution

目的探讨以荧光酶联免疫竞争抑制法(简称Uni CAP法)代替放射性吸附抑制试验(radioallergosorbent test,RAST)测定螨变应原点刺制品生物活性的可行性。方法分别将螨变应原皮肤点刺制品中屋尘螨、粉尘螨点刺液及相应参考品进行系列稀释后,分别与等体积尘螨阳性血清混合,并设阴性对照及100%抑制对照。分别采用γ放射免疫计数器(RAST法)和Immuno CAP 100E全自动过敏原分析仪(Uni CAP法)测定特异性Ig E(s Ig E)浓度,计算点刺样品的生物活性。结果采用Uni CAP法测定结果的RSD值均低于20%。粉尘螨点刺液用两种方法测定的结果基本一致,而屋尘螨点刺液的结果相差较大,但均在标准范围内。结论以Uni CAP法替代RAST法测定点刺液活性具有可行性,可缩短制品的检验时限,并减少了对操作人员及环境的危害。

Objective To investigate the feasibility of fluorescent enzyme-linked immunosorbent inhibition assay instead of radioallergosorbent test （RAST） for determination of bioactivity of mite allergen prick solution in vitro. Methods The Dermatophagoides pteronyssinus and Dermatophagoides farinae prick solutions and the corresponding references were serially diluted, and mixed with an equal volume of mites positive sera respectively, serving negative control and 100% inhibition control. Specific IgE （slgE） concentration was determined by using gamma radioimmunoassay counter and ImmunoCAP 100E automatic allergen analyzer （UniCAP）, based on which the bioactivity of samples was calculated. Results The RSD of determination results by UniCAP was less than 20%. The determination results of Dermatophagoides farinae prick solution by two methods were basically in agareement, while those of Dermatophagoides pteronyssinus varied greatly, all of which were within the range of standard. Conclusion It was feasible to determine the bioactivity of mite allergen prick solution by UnicCAP instead of RAST, by which the test time was shortened, and the harm to operators and the environment were reduced.